NcCyP41, a two domain Neurospora crassa cyclophilin: characterization of its peptidyl-prolyl cis-trans isomerase activity ; isolation and functional analysis of two novel NcCyP41-binding proteins
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
2001
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Freiburg (Breisgau), Univ., Diss., 2001 (Nur beschränkt für den Austausch) |
| Beschreibung: | 93 S. Ill., graph. Darst. : 21 cm |
Internformat
MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV014025670 | ||
| 003 | DE-604 | ||
| 005 | 20030402 | ||
| 007 | t | ||
| 008 | 011120s2001 gw ad|| m||| 00||| eng d | ||
| 016 | 7 | |a 962382965 |2 DE-101 | |
| 035 | |a (OCoLC)248364824 | ||
| 035 | |a (DE-599)BVBBV014025670 | ||
| 040 | |a DE-604 |b ger |e rakddb | ||
| 041 | 0 | |a eng | |
| 044 | |a gw |c DE | ||
| 049 | |a DE-355 | ||
| 100 | 1 | |a Faou, Pierre |d 1971- |e Verfasser |0 (DE-588)123184517 |4 aut | |
| 245 | 1 | 0 | |a NcCyP41, a two domain Neurospora crassa cyclophilin |b characterization of its peptidyl-prolyl cis-trans isomerase activity ; isolation and functional analysis of two novel NcCyP41-binding proteins |c Pierre Faou |
| 264 | 1 | |c 2001 | |
| 300 | |a 93 S. |b Ill., graph. Darst. : 21 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Freiburg (Breisgau), Univ., Diss., 2001 (Nur beschränkt für den Austausch) | ||
| 650 | 4 | |a Neurospora crassa - Cyclophilin | |
| 650 | 0 | 7 | |a Neurospora crassa |0 (DE-588)4171597-4 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Cyclophilin |0 (DE-588)4332417-4 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Neurospora crassa |0 (DE-588)4171597-4 |D s |
| 689 | 0 | 1 | |a Cyclophilin |0 (DE-588)4332417-4 |D s |
| 689 | 0 | |5 DE-604 | |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009604095&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-009604095 | |
Datensatz im Suchindex
| _version_ | 1807322691885072384 |
|---|---|
| adam_text |
TABLE
OF
CONTENTS
1.
INTRODUCTION
.
1
1.1.
THE
FOLDING
PROBLEMS
OF
NEWLY
TRANSLATED
POLYPEPTIDES
.
1
1.2.
ENZYMES
INVOLVED
IN
PROTEIN
FOLDING
.
2
13.
PROLINE
RESIDUES
IN
THE
STRUCTURE-FUNCTION
OF
PROTEINS
.
2
1.4.
ACTIVITY
ASSAYS
.
4
13.
PEPTIDYL-PROLYL
CISLTRANS
ISOMERASES
.
5
13.1.
THE
CYCLOPHILIN
FAMILY
.
5
1.5.2.
THE
FKBP
FAMILY
.
7
13.3.
THE
PARVULIN
FAMILY
.
9
1.6.
IMMUNOSUPPRESSION
BY
CYCLOSPORIN
A,
FK506
AND
RAPAMYCIN.
.
10
1.7.
CELLULAR
FUNCTIONS
OF
LARGE
CYCLOPHILINS
.
10
1.7.1.
CYP40
AND
FKBP52
AS
CONSTITUENTS
OF
THE
MATURE
STEROID
RECEPTOR
COMPLEXES
.
10
1.7.2.
CPR7
AND
HSP90
NEGATIVELY
REGULATE
THE
HEAT
SHOCK
RESPONSE
.
11
1.7.3.
CPR7
AND
HSP90
ARE
INVOLVED
IN
THE
MATURATION
OF
THE
TYROSINE
KINASE
V-SRC
.
12
1.8.
AIM
OF
THE
STUDY
.
12
2.
MATERIALS
AND
METHODS
.
13
2.1.
MATERIALS
AND
EQUIPMENT
.
13
2.1.1.
CHEMICALS
.
13
2.12.
RADIOCHEMICALS
.
14
2.13.
ENZYMES
AND
ANTIBODIES
.
14
2.1.4.
MOLECULAR
BIOLOGY
STANDARDS
AND
KITS
.
14
2.13.
EQUIPMENT
.
14
2.2.
STRAINS,
MEDIA
AND
CULTURES
.
15
2.2.1.
STRAINS
.
15
2.2.1.
J.
ESCHERICHIA
COLI
.
15
2.2.1.2.
SACCHAROMYCES
CEREVISIAE
.
15
2.2.1.3.
NEUROSPORA
CRASSA
.
15
2.2.2.
MEDIA
.
16
2.2.2.1.
E.
COLI-ME&A
.
16
2.2.2.2.
SACCHAROMYCES
CEREVISIAE-MEDIA
.
16
2.2.2.3.
NEUROSPORA
CRASSA-MEDIA
.
17
22.3.
GROWTH
OF
E.
COLI,
S.
CEREVISIAE
AND
N.
CRASSA
.
19
22.3.1.
GROWTH
OF
E.
COLI
CULTURES
.
19
2.2.32.
GROWTH
OF
S.
CEREVISIAE
CULTURES
.
19
22.3.3.
GROWTH
OF
N.
CRASSA
CULTURES
.
19
23.
METHODS
USED
IN
MOLECULAR
BIOLOGY
.
2
0
23.1.
PURIFICATION
OF
PLASMID
DNA
.
20
23.1.1.
ISOLATION
OF
PLASMID
DNA
FROM
E.
COLI
.
20
23.12.
ISOLATION
OF
PLASMID
DNA
FROM
3.
CEREVISIAE
.
20
232.
AGAROSE
GEL
ELECTROPHORESIS
.
20
2.32.1.
CHROMATOGRAPHY
OF
DNA
.
20
2.32.2.
ELUTION
OF
DNA
FROM
AGAROSE
GELS
.
21
233.
ENZYMATIC
MODIFICATION
OF
DNA
.
21
23.3.1.
DIGESTION
OF
DNA
WITH
RESTRICTION
ENDONUCLEASES
.
21
2.3.3.2.
DEPHOSPHORYLATION
OF
DNA
.
21
2.3.3.3.
LIGATION
OF
DNA
FRAGMENTS
.
21
2.3.4.
PREPARATION
AND
TRANSFORMATION
OF
COMPETENT
E.
COLI
AND
S.
CEREVISIAE
.
22
2.3.4.1.
PREPARATION
OF
COMPETENT
E.
COLI
.
22
2.3.4.2.
TRANSFORMATION
OFBACTERIA
.
22
2.3.4.3.
PREPARATION
OF
COMPETENT
S.
CEREVISIAE
.
22
2.3.4.4.
TRANSFORMATION
OF
YEAST
.
22
2.3.5.
SELECTION
OF
CLONES
.23
2.3.6.
PCR
AMPLIFICATION
.
23
2.3.7.
MARATHON
CDNA
LIBRARY
FROM
N.
CRASSA
.
24
2.3.8.
PLASMIDS
.
24
2.3.8.1.
PUC18
.
24
2.3.8.2.
PGEX-5XL
.
24
2.3.8.3.
PET-19B
.
24
2.3.8.4.
PGEM-4Z
.
25
2.3.8.5.
PYES2
.
25
2.4.
METHODS
APPLIED
IN
PROTEIN
BIOCHEMISTRY
.
25
2.4.1.
PURIFICATION
OF
FUSION
PROTEINS
FROM
E.
COLI
.
25
2.4.1.1.
PURIFICATION
OF
HIS-TAG-FUSION
PROTEINS
.
25
2.4.1.2.
PURIFICATION
OF
GST-FUSION
PROTEINS
.
25
2.4.2.
IN
VITRO
TRANSCRIPTION/TRANSLATION
.
26
2.4.2.1.
PREPARATION
OF
N.
CRASSA
LYSATE
.
26
2.4.3.
ESTIMATION
OF
PROTEIN
CONCENTRATION
.
27
2.4.4.
PRECIPITATION
OF
PROTEINS
WITH
DOCA/TCA
.
27
2.4.5.
SDS-POLYACRYLAMIDE
GEL
ELECTROPHORESIS
(SDS-PAGE)
.
27
2.4.6.
NATIVE-POLYACRYLAMIDE
GEL
ELECTROPHORESIS
(NATIVE-PAGE)
.
28
2.4.7.
STAINING
PROTEINS
WITH
COOMASSIE-BLUE
.
28
2.4.8.
AUTORADIOGRAPHY
.
28
2.4.9.
PROTEIN
TRANSFER
ONTO
MEMBRANES
.
29
2.4.9.1.
THE
SEMI-DRY
SYSTEM
.
29
2.4.9.2.
QUANTITATIVE
WESTERN
BLOTTING
.
29
2.4.9.3.
VISUALIZATION
OF
TRANSFERRED
PROTEIN
.
29
2.4.10.
PRODUCTION
OF
POLYCLONAL
ANTISERUM
.
29
2.4.11.
IMMUNOLOGICAL
METHODS
.
30
2.4.11.1.
IMMUNODECORATION
OF
PROTEINS
.
30
2.4.11.2.
COVALENT
COUPLING
OF
ANTIBODIES
TO
PROTEIN-A
SEPHAROSE
.
30
2.4.11.3.
COIMMUNOPRECIPITATION
.
31
2.4.12.
ISOLATION
OF
NCCYP41
ASSOCIATED
PROTEINS
.
31
2.4.13.
DEPICTION
OF
HSP80
IN
THE
LYSATE
.
31
2.4.14.
TWO-DIMENSIONAL
GEL
ELECTROPHORESIS
.
32
2.4.15.
ASSAY
FOR
PEPTIDYL-PROLYL
CIS/TRANS
ISOMERASE
(PPIASE)
ACTIVITY
.
32
3.
RESULTS
.
33
3.1.
N.
CRASSA
CYP41:
STRUCTURE,
CLONING
AND
EXPRESSION.
.
33
3.1.1.
STRUCTURAL
ORGANIZATION
OFN.
CRASSA
CYP41(NCCYP41)
.
33
3.1.2.
CLONING
OFNCCYP41
AND
ITS
DOMAINS
IN
THE
EXPRESSION
VECTOR
PET-19B
.
35
3.1.3.
PURIFICATION
OF(HIS),
0
-NCCYP41
AND
ITS
DOMAINS
FROM
E.
COLI
.
37
3.1.4.
PURIFICATION
OFGST-NCCYP41
.
38
3.2.
PPIASE
ASSAY
OF
PURIFIED
FUSION
PROTEINS
.
39
3.3.
ISOLATION
OF
NCCYP41
BINDING
PROTEINS
.
42
3.3.1.
PRODUCTION
OF
A
N.
CRASSA
CELLULAR
EXTRACT
.
42
3.3.2.
LOADING
AND
ELUTION
OF
INTERACTING
PROTEINS
.
43
3.3.3.
NCCYP41
BINDING
PROTEINS
.
43
3.3.4.
AMINO
ACIDS
SEQUENCES
OF
NCCYP41
ISOLATED
BINDING
PARTNERS
.
45
3.3.5.
CYPBP37
BINDS
DIRECTLY
TO
NCCYP41
.
46
3.4.
CLONING
AND
EXPRESSION
OF
CYPBP37
.
47
3.4.1.
CLONING
OF
A
FULL-LENGTH
CYPBP37
CDNA
.
48
3.4.2.
EXPRESSION
OFCYPBP37
IN
PET-19B
.
49
3.4.3
EXPRESSION
OF
CYPBP37
IN
PGEM-4Z
.
50
3.4.4.
AMINO
ACID
SEQUENCE
OF
CYPBP37
.
51
3.4.5.
PURIFICATION
OF
(HIS)|
0
-CYPBP37
FROM
E.
COLI
.
52
3.5.
PRODUCTION
OF
POLYCLONAL
ANTIBODIES
AGAINST
CYPBP37
.
53
3.6.
CHARACTERIZATION
OF
THE
INTERACTION
BETWEEN
NCCYP41
AND
CYPBP37
.
54
3.6.1,
IN
VITRO
EXPRESSED
CYPBP37
BINDS
TO
(HIS),
0
-NCCYP41
AND
GST-NCCYP41
.
54
3.6.2.
(HIS)
W
-NCCYP41
AND
(HIS)
W
-CYPBP37
FORM
COMPLEXES
IN
NATIVE-PAGE
.
54
3.6.3.
HSP80
AND
CYPBP37
ASSOCIATE
WITH
DISTINCT
DOMAINS
OF
(HIS)
10
-NCCYP4
1
.
57
3.6.4.
BINDING
OF
CYPBP37
TO
THE
PPIASE
DOMAIN
OF
NCCYP4
1
IS
NOT
SENSITIVE
TO
CSA
.
59
3.6.5.
NCCYP4
1
AND
CYPBP37
ALSO
INTERACT
IN
VIVO
.
59
3.6.5.1.
IMMUNOPRECIPITATION
OF
NCCYP41
FROM
A
PROTEIN
EXTRACT
.
60
3.6.5.2.
IMMUNOADSORPTION
OF
CYPBP37
FROM
A
N.
CRASSA
CELL
EXTRACT
.
60
3.7.
TWO-DIMENSIONAL
GEL
ELECTROPHORESIS
OF
NCCYP41
BINDING
PROTEINS.
.
61
3.8.
INFLUENCE
OF
THIAMINE
ON
THE
EXPRESSION
OF
CYPBP37
.
63
3.8.1.
INFLUENCE
OF
THIAMINE
ON
THE
GROWTH
OF
N.
CRASSA
.
63
3.8.2.
EXPRESSION
OF
N.
CRASSA
CYPBP37
IS
REPRESSED
BY
THIAMINE
.
64
3.9.
INFLUENCE
OF
NCCYP41
EXPRESSION
IN
YEAST
.
65
3.9.1.
EXPRESSION
OF
NCCYP41
IN
S.
CEREVISIAE
.
65
3.9.2.
INFLUENCE
OF
NCCYP4
1
ON
THE
GROWTH
OF
S.
CEREVISIAE
THI4
DELETION
MUTANTS.
66
4.
DISCUSSION
.
68
4.1.
STRUCTURAL
ORGANIZATION
OF
NCCYP41
.
68
4.2.
IDENTIFICATION
OF
NCCYP41
BINDING
PROTEINS
.
71
4.3.
CHARACTERIZATION
OF
THE
INTERACTION
BETWEEN
NCCYP41
AND
HSP80
.
73
4.4.
CHARACTERIZATION
OF
THE
BINDING
BETWEEN
NCCYP41
AND
CYPBP37
.
74
4.5.
CHARACTERIZATION
OF
CYPBP37
.
7
6
4.6.
POSSIBLE
ROLE
OF
THE
COMPLEX
CYPBP37-NCCYP41
.
79
5.
SUMMARY
.
80
6.
REFERENCES
.
81 |
| any_adam_object | 1 |
| author | Faou, Pierre 1971- |
| author_GND | (DE-588)123184517 |
| author_facet | Faou, Pierre 1971- |
| author_role | aut |
| author_sort | Faou, Pierre 1971- |
| author_variant | p f pf |
| building | Verbundindex |
| bvnumber | BV014025670 |
| ctrlnum | (OCoLC)248364824 (DE-599)BVBBV014025670 |
| format | Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV014025670 |
| illustrated | Illustrated |
| indexdate | 2024-08-14T01:00:31Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-009604095 |
| oclc_num | 248364824 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | 93 S. Ill., graph. Darst. : 21 cm |
| publishDate | 2001 |
| publishDateSearch | 2001 |
| publishDateSort | 2001 |
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| spelling | Faou, Pierre 1971- Verfasser (DE-588)123184517 aut NcCyP41, a two domain Neurospora crassa cyclophilin characterization of its peptidyl-prolyl cis-trans isomerase activity ; isolation and functional analysis of two novel NcCyP41-binding proteins Pierre Faou 2001 93 S. Ill., graph. Darst. : 21 cm txt rdacontent n rdamedia nc rdacarrier Freiburg (Breisgau), Univ., Diss., 2001 (Nur beschränkt für den Austausch) Neurospora crassa - Cyclophilin Neurospora crassa (DE-588)4171597-4 gnd rswk-swf Cyclophilin (DE-588)4332417-4 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Neurospora crassa (DE-588)4171597-4 s Cyclophilin (DE-588)4332417-4 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009604095&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Faou, Pierre 1971- NcCyP41, a two domain Neurospora crassa cyclophilin characterization of its peptidyl-prolyl cis-trans isomerase activity ; isolation and functional analysis of two novel NcCyP41-binding proteins Neurospora crassa - Cyclophilin Neurospora crassa (DE-588)4171597-4 gnd Cyclophilin (DE-588)4332417-4 gnd |
| subject_GND | (DE-588)4171597-4 (DE-588)4332417-4 (DE-588)4113937-9 |
| title | NcCyP41, a two domain Neurospora crassa cyclophilin characterization of its peptidyl-prolyl cis-trans isomerase activity ; isolation and functional analysis of two novel NcCyP41-binding proteins |
| title_auth | NcCyP41, a two domain Neurospora crassa cyclophilin characterization of its peptidyl-prolyl cis-trans isomerase activity ; isolation and functional analysis of two novel NcCyP41-binding proteins |
| title_exact_search | NcCyP41, a two domain Neurospora crassa cyclophilin characterization of its peptidyl-prolyl cis-trans isomerase activity ; isolation and functional analysis of two novel NcCyP41-binding proteins |
| title_full | NcCyP41, a two domain Neurospora crassa cyclophilin characterization of its peptidyl-prolyl cis-trans isomerase activity ; isolation and functional analysis of two novel NcCyP41-binding proteins Pierre Faou |
| title_fullStr | NcCyP41, a two domain Neurospora crassa cyclophilin characterization of its peptidyl-prolyl cis-trans isomerase activity ; isolation and functional analysis of two novel NcCyP41-binding proteins Pierre Faou |
| title_full_unstemmed | NcCyP41, a two domain Neurospora crassa cyclophilin characterization of its peptidyl-prolyl cis-trans isomerase activity ; isolation and functional analysis of two novel NcCyP41-binding proteins Pierre Faou |
| title_short | NcCyP41, a two domain Neurospora crassa cyclophilin |
| title_sort | nccyp41 a two domain neurospora crassa cyclophilin characterization of its peptidyl prolyl cis trans isomerase activity isolation and functional analysis of two novel nccyp41 binding proteins |
| title_sub | characterization of its peptidyl-prolyl cis-trans isomerase activity ; isolation and functional analysis of two novel NcCyP41-binding proteins |
| topic | Neurospora crassa - Cyclophilin Neurospora crassa (DE-588)4171597-4 gnd Cyclophilin (DE-588)4332417-4 gnd |
| topic_facet | Neurospora crassa - Cyclophilin Neurospora crassa Cyclophilin Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009604095&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT faoupierre nccyp41atwodomainneurosporacrassacyclophilincharacterizationofitspeptidylprolylcistransisomeraseactivityisolationandfunctionalanalysisoftwonovelnccyp41bindingproteins |